CN108490041A - A kind of urease biologic sensor - Google Patents

A kind of urease biologic sensor Download PDF

Info

Publication number
CN108490041A
CN108490041A CN201810166839.0A CN201810166839A CN108490041A CN 108490041 A CN108490041 A CN 108490041A CN 201810166839 A CN201810166839 A CN 201810166839A CN 108490041 A CN108490041 A CN 108490041A
Authority
CN
China
Prior art keywords
solution
platinum
carbon ball
chitosan
biologic sensor
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201810166839.0A
Other languages
Chinese (zh)
Other versions
CN108490041B (en
Inventor
吴丽娜
曹锡忠
周静洁
毛欣
周静珠
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
JIANGSU ACADEMY OF SCIENCE AND TECHNOLOGY FOR INSPECTION AND QUARANTINE
JIANGSU ENTRY-EXIT INSPECTION AND QUARANTINE BUREAU
Original Assignee
JIANGSU ACADEMY OF SCIENCE AND TECHNOLOGY FOR INSPECTION AND QUARANTINE
JIANGSU ENTRY-EXIT INSPECTION AND QUARANTINE BUREAU
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by JIANGSU ACADEMY OF SCIENCE AND TECHNOLOGY FOR INSPECTION AND QUARANTINE, JIANGSU ENTRY-EXIT INSPECTION AND QUARANTINE BUREAU filed Critical JIANGSU ACADEMY OF SCIENCE AND TECHNOLOGY FOR INSPECTION AND QUARANTINE
Priority to CN201810166839.0A priority Critical patent/CN108490041B/en
Publication of CN108490041A publication Critical patent/CN108490041A/en
Application granted granted Critical
Publication of CN108490041B publication Critical patent/CN108490041B/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/308—Electrodes, e.g. test electrodes; Half-cells at least partially made of carbon
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
    • G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
    • G01N27/3278—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction involving nanosized elements, e.g. nanogaps or nanoparticles

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • Electrochemistry (AREA)
  • Analytical Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Engineering & Computer Science (AREA)
  • Nanotechnology (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

The invention discloses a urease biologic sensors, belong to analysis detection field.The sensor is to utilize screen printing electrode, constructs a kind of biosensor based on polyaniline film and chitosan/platinum carbon ball, realizes the quick detection of heavy metal ion.The urease biologic sensor that technical solution of the present invention is prepared has good reproducibility and time saving, laborsaving, cheap, reduction solvent, pollution etc. of the reduction to environment.Sensor is preserved can be placed in 4 DEG C of refrigerator when not used, and the inner potential response of three weeks does not all decline, is remained to keep the 95% of initial potential response after preserving 30 days, is illustrated that chitosan can effectively keep the activity of urase, and can prevent enzyme from leaking.

Description

A kind of urease biologic sensor
The application is：On May 20th, 2016, application No. is：201610340251.3, it is entitled：Based on poly- Aniline modifies the divisional application of the urease biologic sensor of screen printing electrode and its patent of invention of application.
Technical field
The invention belongs to analyze detection field, and in particular to a kind of urease biologic sensor and its application.
Background technology
With popularizing for green consumption idea, ecological textile increasingly becomes the mainstream in market, in recent years, in textile Index one of of the detection of extractable heavy metal as ecological textile is increasingly taken seriously.Heavy metal is main in textile The dyestuff and auxiliary agent used in process, such as various premetallized dyes, phthalein mountain valley with clumps of trees and bamboo structure dyestuff, are consolidated medium fuel Toner, catalyst, fire retardant, post-finishing agent etc. and for soft water hardening, desizing is concise, various in the bleaching processes such as stamp Metal chelating agent.It is absorption of human body that extractable heavy metal, which can be entered by the sweat of human body inside human skin, in textile, Can cause health when heavy metal is accumulated to a certain extent in human organ huge in liver, bone, kidney, the heart and brain Damage.The detection method of extractable heavy metal is mainly GB/T 17593 in the textile of national regulation at present, and which specify spinnings The test method of the various heavies such as arsenic, cadmium, cobalt, chromium, copper, nickel, lead, antimony in fabric.Main analytical instrument to be used is atom Absorption spectrum and inductive coupling plasma emission spectrum.
Although existing analysis method precision is good, accuracy is high, and existing analysis method cannot meet time saving, province Power, cheap, reduction solvent, pollution etc. of the reduction to environment.
Invention content
The present invention be directed to problems of the existing technology to provide a kind of urea based on Polyaniline-modified screen printing electrode Enzyme biologic sensor and its application.
The purpose of the present invention can be achieved through the following technical solutions：
A kind of urease biologic sensor based on Polyaniline-modified screen printing electrode, the sensor are by the following method It is prepared：
(1) screen printing electrode electropolymerization polyaniline：Screen printing electrode is placed in the electricity containing hydrochloric acid and aniline first In polymeric solution, uses cyclic voltammetry to carry out electropolymerization later, obtain PAIN modified electrodes；
(2) platinum/carbon ball nanocomposite：Carbon ball is added in ethylene glycol solution and ultrasonic disperse uniformly obtains carbon Slurry is slowly added to chloroplatinic acid and is uniformly mixing to obtain mixed liquor in carbon slurry, and the mixed liquor is adjusted to alkalinity using alkaline reagent, 100~150 DEG C of reductase 12~6h are warming up to, later by the washing of obtained black product, drying, obtain the nano combined material of platinum/carbon ball Material；
(3) urease biologic sensor：The platinum that chitosan and step (2) are prepared/carbon ball nanocomposite is added To in acetum and being uniformly mixed, chitosan-platinum/carbon ball mixed solution is obtained；By chitosan-platinum/carbon ball mixed solution with Urase solution is uniformly mixed, and obtains urase-chitosan/platinum/carbon ball mixed solution；In the PAIN modifications that step (1) is prepared Electrode surface drop coating urase-chitosan/platinum/carbon ball mixed solution, is prepared urease biologic sensor.Drop coating amount is about 15 μ L/cm2。
A kind of urease biologic sensor preparation method based on Polyaniline-modified screen printing electrode, this method includes following Step：
(1) screen printing electrode electropolymerization polyaniline：Screen printing electrode is placed in the electricity containing hydrochloric acid and aniline first In polymeric solution, uses cyclic voltammetry to carry out electropolymerization later, obtain PAIN modified electrodes；
(2) platinum/carbon ball nanocomposite：Carbon ball is added in ethylene glycol solution and ultrasonic disperse uniformly obtains carbon Slurry is slowly added to chloroplatinic acid and is uniformly mixing to obtain mixed liquor in carbon slurry, and the mixed liquor is adjusted to alkalinity using alkaline reagent, 100~150 DEG C of reductase 12~6h are warming up to, later by the washing of obtained black product, drying, obtain the nano combined material of platinum/carbon ball Material；
(3) urease biologic sensor：The platinum that chitosan and step (2) are prepared/carbon ball nanocomposite is added To in acetum and being uniformly mixed, chitosan-platinum/carbon ball mixed solution is obtained；By chitosan-platinum/carbon ball mixed solution with Urase solution is uniformly mixed, and obtains urase-chitosan/platinum/carbon ball mixed solution；In the PAIN modifications that step (1) is prepared Electrode surface drop coating urase-chitosan/platinum/carbon ball mixed solution, is prepared urease biologic sensor.
In urease biologic sensor described in technical solution of the present invention and preparation method thereof：The electropolymerization process of step (1) It being carried out under conditions of nitrogen protection, cyclic voltammetry scanning range is -0.1V~0.7V, the rate of scanning is 40~ 60mV/s is always scanned 80~120 times.
In urease biologic sensor described in technical solution of the present invention and preparation method thereof：The electropolymerization solution of step (1) A concentration of 0.1~3.5mol/L of middle hydrochloric acid, a concentration of 0.1~3mol/L of aniline.
In urease biologic sensor described in technical solution of the present invention and preparation method thereof：The carbon ball of step (2) and chlorine platinum The mass ratio of acid is 1~3：1.
In urease biologic sensor described in technical solution of the present invention and preparation method thereof：The alkalinity of step (2) refers to mixing The pH value for closing liquid is 9~11.
In urease biologic sensor described in technical solution of the present invention and preparation method thereof：Chitosan-platinum of step (3)/ The mass fraction of chitosan is 0.1~1% in carbon ball mixed solution, the mass fraction of platinum/carbon ball nanocomposite is 0.5~ 1.5%.
In urease biologic sensor described in technical solution of the present invention and preparation method thereof：The urase solution of step (3) is dense It is 5~15mg/ml to spend, and the volume ratio of chitosan-platinum/carbon ball mixed solution and urase solution is 1~5：1~5.
In urease biologic sensor described in technical solution of the present invention and preparation method thereof：The drop coating amount of step (3) is 10 ~30 μ L/cm2。
Application of the aforementioned PAIN modified electrodes in measuring pH value of solution；It is preferred that the inhibition time measured be >= 20min, response time >=2min.
Application of the aforementioned urease biologic sensor in detecting heavy metal ion；It is preferred that being detected in urea liquid Hg2+And Cd2+In application；Hg in textile is detected more preferably in urea liquid2+And Cd2+Application.The inhibition time of measurement For >=20min, response time >=2min；It is preferred that the concentration of urea liquid≤selection 40mmol/L.
Beneficial effects of the present invention：
The urease biologic sensor that technical solution of the present invention is prepared have good reproducibility and it is time saving, laborsaving, cheap, It reduces solvent, reduce the pollution etc. to environment.Sensor is preserved can be placed in 4 DEG C of refrigerator when not used, in three weeks Potential response does not all decline, remains to keep the 95% of initial potential response after preserving 30 days, illustrates that chitosan can be effectively The activity of urase is kept, and can prevent enzyme from leaking.
Description of the drawings
Fig. 1 is the voltammogram that 1 aniline monomer of embodiment powers on 100 gained of polymerisation loop in screen printing electrode.
Before and after Fig. 2 is aniline polymerization, the cycle of screen printing electrode is bent over the desk figure.
Fig. 3 is the relational graph of the potential response and solution ph of PAIN modified electrodes.
Fig. 4 is that urease biologic sensor is placed in the potential response of urea liquid and changes with time figure.
Fig. 5 is in Hg2+Or Cd2+In the presence of, inhibiting rate, which changes with time, sees Fig. 5
Fig. 6 is that urease biologic sensor is placed in the solution containing urea, urea concentration and potential change figure.
Fig. 7 is that urease biologic sensor is placed in the solution containing urea, the relational graph of inhibiting rate and heavy metal concentration.
Specific implementation mode
With reference to embodiment, the present invention will be further described, and but the scope of the present invention is not limited thereto：
Carbon ball described in the embodiment of the present invention can be that commercial product is either prepared with the following method：With first Aldehyde-resorcinol is precursor, is usedMethod, 0.1mL ammonium hydroxide (25w%) are added to 8mL ethyl alcohol and 20mL deionized waters Mixed solution in, stir 1h after, be added 0.2g resorcinols, stir to being completely dissolved, it is molten that 0.284mL formaldehyde is then added dropwise again Liquid (37w%), mixed liquor stir for 24 hours at 30 DEG C, then move to mixed liquor in water heating kettle, are reacted at 100 DEG C for 24 hours, mixture It is centrifuged, dry 48h at 100 DEG C.Next, carry out charing process is heated to 350 with the heating rate of 1 DEG C/min DEG C, 2h is kept, is then heated to 600 DEG C again with the heating rate of 1 DEG C/min, calcining 4h is retained, then naturally cools to room Carbon ball is prepared in temperature.
Embodiment 1
Screen printing electrode electropolymerization polyaniline：Screen printing electrode is placed in the electropolymerization containing hydrochloric acid and aniline first In solution, wherein the molar concentration of HC1 is 1mol/L, and the molar concentration of aniline is 0.5mol/L；Cyclic voltammetry is used later Electropolymerization is carried out, cyclic voltammetry scanning range is -0.1V~0.7V, sweep speed 50mV/s, is scanned 100 times in total.It is real Electropolymerization solution need to lead to nitrogen l0min before testing beginning, and entire electropolymerization process is completed under a nitrogen atmosphere.After the completion of electropolymerization, use 1mol/L hydrochloric acid and secondary water are rinsed electrode, dry, and PAIN modified electrodes are made；
Platinum/carbon ball nanocomposite：80mg carbon balls are added in ethylene glycol solution and ultrasonic disperse uniformly obtains carbon Slurry is slowly added to 42mg chloroplatinic acids in carbon slurry and is uniformly mixing to obtain mixed liquor, the pH of mixed is adjusted using NaOH solution To 10, it is warming up to 130 DEG C of reduction 3h and obtains platinum/carbon ball nanocomposite later by the washing of obtained black product, drying；
Urease biologic sensor：Platinum/carbon ball nanocomposite that chitosan and step (2) are prepared is added to vinegar It in acid solution and is uniformly mixed, obtains chitosan-platinum/carbon ball mixed solution, the mass fraction of chitosan is in the mixed liquor 0.5%, the mass fraction of platinum/carbon ball nanocomposite is 1%；It is 1 by volume ratio：1 chitosan-platinum/carbon ball mixing is molten Liquid is uniformly mixed with a concentration of 10mg/ml urases solution, obtains urase-chitosan/platinum/carbon ball mixed solution；It is made in step (1) Standby obtained 15 μ L/cm of PAIN modified electrodes surface drop coating2Urase-chitosan/platinum/carbon ball mixed solution, is prepared urase Biosensor.
Embodiment 2
Screen printing electrode electropolymerization polyaniline：Screen printing electrode is placed in the electropolymerization containing hydrochloric acid and aniline first In solution, wherein the molar concentration of HC1 is 2mol/L, and the molar concentration of aniline is 1mol/L；Later use cyclic voltammetry into Row electropolymerization, cyclic voltammetry scanning range are -0.1V~0.7V, sweep speed 40mV/s, are scanned 80 times in total.Experiment is opened Electropolymerization solution need to lead to nitrogen l0min before beginning, and entire electropolymerization process is completed under a nitrogen atmosphere.After the completion of electropolymerization, 1mol/L is used Hydrochloric acid and secondary water are rinsed electrode, dry, and PAIN modified electrodes are made；
Platinum/carbon ball nanocomposite：50mg carbon balls are added in ethylene glycol solution and ultrasonic disperse uniformly obtains carbon Slurry is slowly added to 42mg chloroplatinic acids in carbon slurry and is uniformly mixing to obtain mixed liquor, the pH of mixed is adjusted using NaOH solution To 9, it is warming up to 100 DEG C of reduction 6h and obtains platinum/carbon ball nanocomposite later by the washing of obtained black product, drying；
Urease biologic sensor：Platinum/carbon ball nanocomposite that chitosan and step (2) are prepared is added to vinegar It in acid solution and is uniformly mixed, obtains chitosan-platinum/carbon ball mixed solution, the mass fraction of chitosan is in the mixed liquor 1%, the mass fraction of platinum/carbon ball nanocomposite is 1.5%；It is 1 by volume ratio：3 chitosan-platinum/carbon ball mixing is molten Liquid is uniformly mixed with a concentration of 5mg/ml urases solution, obtains urase-chitosan/platinum/carbon ball mixed solution；It is made in step (1) Standby obtained 10 μ L/cm of PAIN modified electrodes surface drop coating2Urase-chitosan/platinum/carbon ball mixed solution, is prepared urase Biosensor.
Embodiment 3
Screen printing electrode electropolymerization polyaniline：Screen printing electrode is placed in the electropolymerization containing hydrochloric acid and aniline first In solution, wherein the molar concentration of HC1 is 3mol/L, and the molar concentration of aniline is 1.5mol/L；Cyclic voltammetry is used later Electropolymerization is carried out, cyclic voltammetry scanning range is -0.1V~0.7V, sweep speed 60mV/s, is scanned 120 times in total.It is real Electropolymerization solution need to lead to nitrogen l0min before testing beginning, and entire electropolymerization process is completed under a nitrogen atmosphere.After the completion of electropolymerization, use 1mol/L hydrochloric acid and secondary water are rinsed electrode, dry, and PAIN modified electrodes are made；
Platinum/carbon ball nanocomposite：120mg carbon balls are added in ethylene glycol solution and ultrasonic disperse uniformly obtains carbon Slurry is slowly added to 42mg chloroplatinic acids in carbon slurry and is uniformly mixing to obtain mixed liquor, the pH of mixed is adjusted using NaOH solution To 11,150 DEG C of reductase 12 h are warming up to, later by the washing of obtained black product, drying, obtain platinum/carbon ball nanocomposite；
Urease biologic sensor：Platinum/carbon ball nanocomposite that chitosan and step (2) are prepared is added to vinegar It in acid solution and is uniformly mixed, obtains chitosan-platinum/carbon ball mixed solution, the mass fraction of chitosan is in the mixed liquor 0.4%, the mass fraction of platinum/carbon ball nanocomposite is 1.5%；It is 3 by volume ratio：1 chitosan-platinum/carbon ball mixing Solution is uniformly mixed with a concentration of 15mg/ml urases solution, obtains urase-chitosan/platinum/carbon ball mixed solution；In step (1) The 30 μ L/cm of PAIN modified electrodes surface drop coating being prepared2Urase-chitosan/platinum/carbon ball mixed solution, is prepared urea Enzyme biologic sensor.
Performance detection
The preparation of 1 PAIN modified electrodes
Fig. 1 is the voltammogram that 1 aniline monomer of embodiment powers on 100 gained of polymerisation loop in screen printing electrode, is followed Ring voltammogram shows that its electrode process has invertibity.Spike potential is aoxidized respectively in 0.22V, and reduction spike potential is in 0.08V.The peak The process of radical cation is oxidized to for the aniline of protonation.With the increase of cycle-index, peak current increases, this is because Aniline is once in electrode surface polymerization film formation, it may occur that self-catalyzed reaction, while the film thickness of polymer also gradually increases.It prepares Obtained polyaniline film modified electrode is blue-green.Before and after aniline polymerization, the cycle of screen printing electrode figure of bending over the desk is shown in Fig. 2, bent Line a is before electropolymerization starts, and the cyclic voltammogram of screen printing electrode, curve b is after the completion of electropolymerization, and polyaniline film covers The screen printing electrode of lid recycles figure of bending over the desk.Figure it is seen that after electropolymerization, the Polyaniline-modified being prepared is electric Pole has a pair of apparent redox peaks, peak current to significantly increase, and illustrates that polyaniline film cathode is successfully prepared.
The pH of 2 PAIN modified electrodes is responded
PAIN modified electrodes have response to the current potential of solution, the pH value variation for the solution that can be surveyed.The potential response of electrode It is obtained with the relationship of solution ph by detecting the open circuit potential of itself and Ag/AgC1 reference electrodes.Different pH value solution, can be with Different potential values is obtained, the results are shown in Figure 3.The response range of this Polyaniline-modified screen printing electrode be pH 1~ PH12, shows preferable linear relationship, linearly dependent coefficient 0.999, and slope is：60.8mV/pH(T：25 DEG C), see Fig. 3. From the figure 3, it may be seen that polyaniline modified electrode has good potential response.
3 response times and the selection for inhibiting the time
The hydrolysis of urease biologic sensor catalyzing urea needs certain response time, should ensure that when being tested identical Response time.Urease biologic sensor is placed in urea liquid, its potential response is tested and changes with time, concrete outcome See Fig. 4, from fig. 4, it can be seen that potential response increase at any time and increase, tend towards stability after the 2 minutes.Therefore in detection electricity When position, it is 2min to select the response time i.e. time of enzymatic reacting of electrode.
In the presence of having heavy metal ion in the solution, the activity of urase can be suppressed.With the growth of time, Inhibiting rate can increase, and inhibiting rate reaches balance within a certain period of time.Sensor is placed in containing Hg2+Or Cd2+With the solution of urea In, inhibiting rate, which changes with time, sees Fig. 5, it can be seen that from 0-20min, with the growth of time, inhibiting rate increases, After 20min, inhibiting rate reaches a platform, therefore when carrying out heavy metal analysis, and the inhibition time selected is 20min.
The selection of 4 urea concentrations
The urease biologic sensor that embodiment 1 is prepared is placed in the solution containing urea, urase can catalyzing urea Hydrolysis, generate CO2And NH3, cause the potential change of solution.The concentration of urea liquid can influence the degree of potential change, urea Concentration is shown in Fig. 6 from the relationship of 10-100mmol/L and potential change.From fig. 6, it can be seen that urea concentration is from 10 to 40mmol/L When, potential difference is directly proportional to urea concentration, urea concentration from 50mmol/L to 100mmol/L potential difference with urea concentration Variation, speedup slows down, this is because the fixed enzyme of electrode surface, the urea of catalysis Finite Concentration is only capable of, when urea concentration mistake Gao Shi, the catalytic capability of enzyme can tend to be saturated.According to Fig.6, the urea liquid of selection≤40mmol/L, the most properly.
5 urease biologic sensors are used for the detection of heavy metal ion
The urease biologic sensor that embodiment 1 is prepared is placed in the urea liquid of 40mmol/L, it is anti-to measure enzymatic Potential change Δ E caused by answering.The Hg of various concentration is separately added into urea liquid2+(10、50、100、500、1000、2000 μ g/L) and Cd2+(50,100,500,1000,2000,5000 μ g/L) solution, after twenty minutes, the current potential after test inhibition calculates The potential change Δ E' of enzymatic reaction after inhibition.Heavy metal to the inhibiting rate of urease activity be equal to [(Δ E- Δ E')/Δ E] × 100%.
The relationship of inhibiting rate and heavy metal concentration is as shown in fig. 7, it can be seen from figure 7 that Hg2+And Cd2+Inhibiting rate Increase with the increase of concentration, the inhibiting rate of enzymatic activity and the negative logarithm of concentration are in good linear relationship, linear correlation system Number is in 0.99 or more, Hg2+The range of linearity is 10-2000 μ g/L, Cd2+The range of linearity be 500-5000 μ g/L, calculate detection It is limited to Hg2+3.89 μ g/L, Cd2+5.41 μ g/L (are calculated) by inhibiting rate 10%.
The reproducibility and stability of 6 urease biologic sensors
The 500 μ g/L Hg of urease biologic sensor pair for selecting embodiment 1 to be prepared2+With 500 μ g/L Cd2+Detection Relative standard deviation is respectively 7.2%, 8.9%.Show that the sensor has good reproducibility.Sensor can set when not used It is preserved in 4 DEG C of refrigerator, the inner potential response of three weeks does not all decline, remains to keep initial potential response after preserving 30 days 95%, illustrate that chitosan can effectively keep the activity of urase, and can prevent enzyme from leaking.
Detection of 7 sensors to textile actual sample
The preparation of textile samples solution：Textile samples first shred into 5mm × 5mm fragments, weigh 2g, and 80mL acid is added Property sweat, be put into thermostatic control oscillator vibration and vibrate after sixty minutes, it is cooling for use.Acidic sweat is according to GB/T17593.2-2007 Requirement prepare.
The urease biologic sensor that embodiment 1 is prepared is selected to carry out mark-on reclaims in textile samples extracting solution, Calculate its rate of recovery.Particular content is shown in Table 1, as it can be seen from table 1 the rate of recovery range of sample is between 80%-120%, it can For the detection of actual sample.
The recovery of standard addition of 1 actual sample of table

Claims (5)

1. a kind of urease biologic sensor, it is characterised in that：The sensor is to be prepared via a method which to obtain：
(1) screen printing electrode electropolymerization polyaniline：Screen printing electrode is placed in the electropolymerization containing hydrochloric acid and aniline first In solution, uses cyclic voltammetry to carry out electropolymerization later, obtain PANI modified electrodes；
(2) platinum/carbon ball nanocomposite：Carbon ball is added in ethylene glycol solution and ultrasonic disperse uniformly obtains carbon slurry, It is slowly added to chloroplatinic acid in carbon slurry and is uniformly mixing to obtain mixed liquor, which is adjusted to alkalinity, heating using alkaline reagent To 100~150 DEG C of reductase 12~6h, later by the washing of obtained black product, drying, platinum/carbon ball nanocomposite is obtained；
(3) urease biologic sensor：Platinum/carbon ball nanocomposite that chitosan and step (2) are prepared is added to vinegar It in acid solution and is uniformly mixed, obtains chitosan-platinum/carbon ball mixed solution；By chitosan-platinum/carbon ball mixed solution and urase Solution is uniformly mixed, and obtains urase-chitosan/platinum/carbon ball mixed solution；In the PANI modified electrodes that step (1) is prepared Surface liquid-transfering gun drop coating urase-chitosan/platinum/carbon ball mixed solution, is prepared urease biologic sensor；
Wherein：The electropolymerization process of step (1) carries out under conditions of nitrogen protection, cyclic voltammetry scanning range be- The rate of 0.1V~0.7V, scanning are 40~60mV/s, are always scanned 80~120 times；The quality of the carbon ball and chloroplatinic acid of step (2) Than being 1~3：1.
2. urease biologic sensor according to claim 1, it is characterised in that：Hydrochloric acid in the electropolymerization solution of step (1) A concentration of 0.1~3.5mol/L, a concentration of 0.1~3mol/L of aniline.
3. urease biologic sensor according to claim 1, it is characterised in that：The alkalinity of step (2) refers to mixed liquor PH value is 9~11.
4. urease biologic sensor according to claim 1, it is characterised in that：Chitosan-platinum/carbon ball of step (3) is mixed The mass fraction for closing chitosan in solution is 0.1~1%, and the mass fraction of platinum/carbon ball nanocomposite is 0.5~1.5%.
5. urease biologic sensor according to claim 1, it is characterised in that：The urase solution concentration of step (3) be 5~ The volume ratio of 15mg/ml, chitosan-platinum/carbon ball mixed solution and urase solution is 1~5：1~5, drop coating amount is about 10~30 μ L/cm2。
CN201810166839.0A 2016-05-20 2016-05-20 A kind of urease biologic sensor Expired - Fee Related CN108490041B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201810166839.0A CN108490041B (en) 2016-05-20 2016-05-20 A kind of urease biologic sensor

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201810166839.0A CN108490041B (en) 2016-05-20 2016-05-20 A kind of urease biologic sensor
CN201610340251.3A CN106018509B (en) 2016-05-20 2016-05-20 Urease biologic sensor based on Polyaniline-modified screen printing electrode and its application

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
CN201610340251.3A Division CN106018509B (en) 2016-05-20 2016-05-20 Urease biologic sensor based on Polyaniline-modified screen printing electrode and its application

Publications (2)

Publication Number Publication Date
CN108490041A true CN108490041A (en) 2018-09-04
CN108490041B CN108490041B (en) 2019-02-19

Family

ID=57096709

Family Applications (4)

Application Number Title Priority Date Filing Date
CN201810168856.8A Expired - Fee Related CN108414587B (en) 2016-05-20 2016-05-20 A kind of preparation method of urease biologic sensor
CN201810167584.XA Expired - Fee Related CN108490042B (en) 2016-05-20 2016-05-20 A kind of purposes of urease biologic sensor
CN201810166839.0A Expired - Fee Related CN108490041B (en) 2016-05-20 2016-05-20 A kind of urease biologic sensor
CN201610340251.3A Expired - Fee Related CN106018509B (en) 2016-05-20 2016-05-20 Urease biologic sensor based on Polyaniline-modified screen printing electrode and its application

Family Applications Before (2)

Application Number Title Priority Date Filing Date
CN201810168856.8A Expired - Fee Related CN108414587B (en) 2016-05-20 2016-05-20 A kind of preparation method of urease biologic sensor
CN201810167584.XA Expired - Fee Related CN108490042B (en) 2016-05-20 2016-05-20 A kind of purposes of urease biologic sensor

Family Applications After (1)

Application Number Title Priority Date Filing Date
CN201610340251.3A Expired - Fee Related CN106018509B (en) 2016-05-20 2016-05-20 Urease biologic sensor based on Polyaniline-modified screen printing electrode and its application

Country Status (1)

Country Link
CN (4) CN108414587B (en)

Families Citing this family (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108414587B (en) * 2016-05-20 2019-01-04 江苏出入境检验检疫局工业产品检测中心 A kind of preparation method of urease biologic sensor
CN109880882B (en) * 2019-03-29 2022-03-25 山东时进检测服务有限公司 Method for detecting lead pollution in marine food
CN109880881B (en) * 2019-03-29 2022-03-25 山东时进检测服务有限公司 Method for detecting cadmium pollution in marine food
CN111020004B (en) * 2019-12-28 2022-09-30 哈尔滨工业大学 Preparation method of urea sensor with Janus structure artificial cell model
CN113433087A (en) * 2021-06-22 2021-09-24 中南大学 Rapid detection method for urea concentration, detection sensor and application
CN113484388B (en) * 2021-06-30 2022-04-12 山东大学 Method for screening helicobacter pylori urease inhibitor
CN114605046B (en) * 2022-01-21 2023-06-16 西安建筑科技大学 Chitosan reinforced enzyme induced carbonate precipitation curing agent and application method thereof
CN114965634A (en) * 2022-04-14 2022-08-30 深圳可孚生物科技有限公司 Preparation method of silk-screen bioelectrochemical sensor
CN115266865B (en) * 2022-07-29 2024-04-16 北京大学 Method for improving stability of electrochemical sensor

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101251506A (en) * 2008-03-15 2008-08-27 西北师范大学 Method for manufacturing conductive polyaniline/cellulose composite biosensor
CN103421878A (en) * 2013-07-26 2013-12-04 江苏大学 Preparation of polyphenol oxidase biosensor and detection of polyphenol oxidase biosensor to pesticide residues
CN104034786A (en) * 2014-06-26 2014-09-10 华东理工大学 Preparation of superoxide anion sensor based on noble metal alloy/carbon composite material
US20150198556A1 (en) * 2014-01-10 2015-07-16 National Central University Sensing electrode of enzyme-based sensor and method for manufacturing the same

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102628830B (en) * 2012-04-23 2014-06-11 南京工业大学 Full-solid urase electrode and preparation method thereof
CN104198554B (en) * 2014-09-03 2016-06-15 海南大学 A kind of working electrode and preparation method thereof, biosensor
CN104914150B (en) * 2015-06-15 2018-11-20 上海交通大学 A kind of pH detecting electrode based on graphene/polyaniline composite membrane
CN105548316B (en) * 2015-12-31 2018-04-03 中国农业大学 Heavy metal-polluted soil detector based on urease biologic sensor
CN108414587B (en) * 2016-05-20 2019-01-04 江苏出入境检验检疫局工业产品检测中心 A kind of preparation method of urease biologic sensor

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101251506A (en) * 2008-03-15 2008-08-27 西北师范大学 Method for manufacturing conductive polyaniline/cellulose composite biosensor
CN103421878A (en) * 2013-07-26 2013-12-04 江苏大学 Preparation of polyphenol oxidase biosensor and detection of polyphenol oxidase biosensor to pesticide residues
US20150198556A1 (en) * 2014-01-10 2015-07-16 National Central University Sensing electrode of enzyme-based sensor and method for manufacturing the same
CN104034786A (en) * 2014-06-26 2014-09-10 华东理工大学 Preparation of superoxide anion sensor based on noble metal alloy/carbon composite material

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
CHARLES LUHANA: "A novel enzymatic glucose sensor based on Pt nanoparticles-decorated hollow carbon spheres-modified glassy carbon electrode", 《JOURNAL OF NANOPARTICLE RESEARCH》 *
YUNG-CHIEN LUO 等: "Urea biosensor based on PANi(urease)-Nafion®/Au composite electrode", 《BIOSENSORS & BIOELECTRONICS》 *

Also Published As

Publication number Publication date
CN106018509B (en) 2018-07-31
CN106018509A (en) 2016-10-12
CN108490041B (en) 2019-02-19
CN108414587A (en) 2018-08-17
CN108490042A (en) 2018-09-04
CN108414587B (en) 2019-01-04
CN108490042B (en) 2019-03-12

Similar Documents

Publication Publication Date Title
CN106018509B (en) Urease biologic sensor based on Polyaniline-modified screen printing electrode and its application
Li et al. A robust electrochemical sensing of molecularly imprinted polymer prepared by using bifunctional monomer and its application in detection of cypermethrin
CN103926294B (en) Preparation and application of CS/IL-GR modified bovine serum albumin molecular imprinting electrode
CN104181217A (en) Construction and application of impedance type electrochemical sensor based on molecularly imprinted polymer with magnetic surface
CN105259243A (en) Preparing method of molybdenum sulfide compound and application of molybdenum sulfide compound in detecting hexavalent chromium in water
CN104764775A (en) Method for preparation of triazole molecularly imprinted electrochemical sensor based on electropolymerized OPD/OAP composite membrane and application thereof
CN111440286A (en) Preparation method and application of chiral Co-MOF/CoSR/PU core-shell composite material
CN110057878A (en) A kind of molecular imprinting electrochemical detection method of ketamine drugs
CN107300582A (en) Antibiotic detection means and its manufacture method, detection method based on microsensor
CN109254049A (en) A kind of preparation method and application of ampicillin sensor
CN109254052A (en) A kind of preparation method and application of organophosphorus pesticide electrochemical luminescence sensor
CN106248609A (en) A kind of ultraviolet spectrophotometer measures the method for hexafluorophosphoric acid lithium content in lithium-ion battery electrolytes
CN110763776B (en) LC-HG-AFS detection method of thioarsenate
CN112694072A (en) Fe-Ni metal nitride composite material, preparation method thereof and application thereof in glucose sensor
CN109254060A (en) A kind of preparation method and application of clenbuterol hydrochloride electrochemical sensing electrode
CN109254059A (en) A kind of preparation method and application of tetracycline antibiotics molecular imprinting electrochemical sensor
CN109307698A (en) A kind of preparation method and application for the iron cobalt nitride sensing electrode detecting organo-chlorine pesticide
CN109307695A (en) A kind of preparation method and application of Spanon Electrochemiluminescsensor sensor
CN109212008B (en) Preparation method and determination method of electrochemical sensor for detecting neonicotinoid pesticides
CN109254062A (en) A kind of preparation method and application of macrolide antibiotics molecular engram electrochemical sensor
WO2020224490A1 (en) Nickel foam self-supporting electrode material, preparation method therefor, and use thereof in detection of hydrogen phosphate ion
CN109192355B (en) Solid conductive gel suitable for electrochemical system and preparation method thereof
CN109254057A (en) A kind of preparation method and application of chrysanthemum ester insecticide electrochemical sensing electrode
CN109254050A (en) A kind of preparation method and application of clenbuterol hydrochloride Electrochemiluminescsensor sensor
CN109254054A (en) A kind of preparation method and application of the Spanon sensor based on cobalt-based nitride nano array

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
CB03 Change of inventor or designer information
CB03 Change of inventor or designer information

Inventor after: Wu Mengbi

Inventor after: Cao Xizhong

Inventor after: Wu Lina

Inventor after: Dong Shaowei

Inventor after: Zhou Jingzhu

Inventor before: Wu Lina

Inventor before: Cao Xizhong

Inventor before: Zhou Jingjie

Inventor before: Mao Xin

Inventor before: Zhou Jingzhu

GR01 Patent grant
GR01 Patent grant
CF01 Termination of patent right due to non-payment of annual fee
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20190219

Termination date: 20210520